Vaccine composition and method of using the same

ABSTRACT

The present invention relates to a stable compacted, compressed or hard tableted injectable composition, including a vaccine composition comprising at least one freeze dried antigenic component and a dissolution aid. A package containing the above injectable composition and method to facilitate immunizing a subject against a disease comprising the steps of first dissolving the compacted, compressed or hard tableted vaccine composition in a package with a diluent to form a vaccine solution, and administering the resulting vaccine solution in an amount effective for immunizing is also provided.

This application is a continuation-in-part and claims priority of U.S.Ser. No. 09/642,913, filed Aug. 21, 2000, which claims the benefit ofU.S. Provisional Application No. 60/150,514, filed Aug. 24, 1999. Thecontents of the above-identified U.S. provisional application are herebyincorporated by reference into this application.

Through this application, various references are identified by authorsand full citations. Disclosure of these publications in their entiretiesare hereby incorporated by reference into this application to more fullydescribe the state of the art to which this invention pertains.

FIELD OF THE INVENTION

The present invention relates to a stable compacted, compressed ortableted vaccine composition having a dense solid form, comprising oneor more freeze dried (lyophilized) antigenic components and adissolution aid. This stable dense vaccine composition retains all ofthe advantageous properties of the lyophilized components thereof,including titer stability and solubility while further providingcomplete and rapid dissolution in a diluent. Moreover, the vaccinecomposition of the present invention avoids the disadvantages of priorknown lyophilized preparations. A method for immunizing a subject usingthe stable vaccine composition to form a vaccine solution is alsoprovided.

BACKGROUND OF THE INVENTION

It is well-known, that biological materials in solutions such asvaccines are susceptible to varying influences including heat, oxidizingreagents, salts, pH, light, and proteolitic enzymes.

Several methods are known for reducing these detrimental effects ingeneral and more specifically to improve the stability of a vaccineespecially during storage. For example, storage below 0° C., and as lowas −70° C., in a freezer is a well-known method. At even lowertemperatures, e.g. in liquid nitrogen, many biological materialsincluding living cells can successfully be stored for many years.However, such methods are not always convenient in those situations, forexample, that involve the innoculation of free-ranging livestock.

Lyophilizing or freeze drying is another known way of conserving livecells and viruses for use as vaccines. During freeze drying, thesolution containing the biological material is first frozen and thewater is then evaporated by sublimation, usually under high vacuum andsub-zero temperatures. Previous approaches have used freeze drying orother techniques to formulate viral vaccines but still pose otherdifficulties with respect to preparation and administration of a stablefinal dosage form.

For example, U.S. Pat. No. 4,251,509 (Hanson and Abegunde) discloses astable particulate viral vaccine intended to be orally administered tofree ranging livestock in a dry state. However, the disclosed dosageform is not freeze dried but prepared by concentrating and extruding apaste dried to form pellets.

Antioxidants, the selection of which will depend on the particularvirus, are required to promote thermostability. Such a formulation maybe more complicated to prepare and is not particularly well-suited forthose instances when it is desired to dissolve the vaccine beforeimmunization.

U.S. Pat. Nos. 3,458,621 and 3,608,030 (Tint) disclose the use of freezedried virus preparations to prepare a tablet for oral administration forthe immunization of the intestinal tract. However, the tablets preparedare for oral administration and are provided with an enteric coating todelay disintegration. During in-vitro testing they did not rapidlydisintegrate and were found to disintegrate only “within 25 minutes”with simulated intestinal fluid. Moreover, the time to disintegrate wasmeasured in simulated intestinal fluid thereby discounting effects ofthe enteric coating. In addition, Tint further emphasizes that unlessthe tablets are “press coated” they will lose their titer as shown by acomparison between press coated and non-press coated tablets. Tint notesthat the non-press coated tablets “not only failed to elicit an antibodyresponse in all the antibody-negative individuals, but, in addition, themagnitude of the titer rise was significantly smaller.” (See column 4,line 40).

PCT Publication No. WO 99/21579 (Seager, et al.) assigned to the R. P.Scherer Corp. discloses a “fast” dispersing composition for a veterinaryvaccine such as against New Castle disease that is freeze dried and“loosely compacted.” The dosage form is disclosed as an “open matrixnetwork”, such as a “solid foam” referenced from U.S. Pat. No. 4,371,516(counterpart to UK Patent No. 1,548,022), as opposed to a compressedform or hard tableting. Moreover, the vaccines are directed to oraladministration and targeted towards retention at mucosal tissue.Adjuvants serve to provide sufficient residence time for absorptionthereon. The disclosed vaccine formulation does not provide forpreparation of a liquid dosage for later immunizing, nor a means toreadily form a stable, measured vaccine solution for administration as aliquid dose, nor for providing a stable compressed or hard tabletedlyophilisate to facilitate later administration thereof.

U.S. Pat. No. 5,587,180 (Allen, Jr. et al) describes a process formaking a particulate support matrix for a rapidly dissolving tablet. Theprocess teaches away from freeze drying and uses standard spray-dryingtechniques. The particulate support matrix is suitable for dosageadministration when placed into the oral cavity. Moreover, no stablevaccine formulation is provided as a vaccine solution in a liquid dosageform.

U.S. Pat. No. 5,336,666 (Neway et al.) discloses a freeze dried liquidvaccine that may form a tablet to be reconstituted in liquid form.However, the vaccine is limited to a polar glycopeptide of a particularbacterium and does not provide for complete or rapid dissolution.

Although freeze drying of biological material can be performed accordingto lyophilizing procedures well known to one skilled in the art toprovide a stable vaccine preparation, the titer of a live virus at theend of lyophilization is typically not the same as it was for thesolution before the lyophilization process. In general it is notpossible to conduct titration before lyophilization as the solution isnot stable until it is freeze dried. In addition, the titer will changeunpredictably during lyophilization. Consequently, an estimate for theinitial titer based on experience is only validated by titration afterfreeze drying. As a result of all of the above, it is almost impossibleto achieve a defined accurate target titer.

If lyophilization has been carried out with the lyophilisate alreadycontained in vials, reworking of the batch is typically not possible,and in some cases a whole batch must be discarded if it is not up tospecification.

U.S. Pat. No. 5,897,852 (Wilderbeek, et al.) attempts to solve such aproblem using different “freeze dried bodies” with “lyospheres” to makeup for shortfalls in the titer of a lyophilized cake. However, eachlyosphere has its own titer thereby necessitating the titration ofmultiple bodies to arrive at a desired titer. Even in the best case,this method does not achieve the exact required titer as it is only anapproximation due to the use of combined bodies to achieve the targettiter. Furthermore, the production of lyospheres is relatively difficultcompared with the more straightforward freeze drying of solutions toproduce a cake or powder. Additionally, a special matrix is oftenrequired to prevent the lyosphere material from being pulverized afterdrying. In general, the method requires the preparation of separatesolutions each having a different titer, additives and adjuvants. Themethod does not solve the problem that the titer for a batch of livevirus is rarely homogenous, and varies from one vial to the next,particularly over the areas of the cold plates in the lyophiliser.Therefore, determination of the number of lyospheres needed per vial isalways an approximation.

Freeze drying may also be useful for vaccines comprising more than oneimmunogenic component. For example, EP 290197A discloses a freeze driedtetravalent vaccine. The procedure discloses the mixing and subsequentfreeze drying of four live virus vaccine components.

A disadvantage to current freeze drying techniques for vaccinepreparation is that the process is very complex, having many variables,rendering it notoriously difficult to perform in a reproducible mannerto achieve acceptable product and dosage uniformity. This is especiallyproblematic for veterinary vaccines where a large number of doses arefreeze dried in one vial. The problem is less common for human vaccinepreparation, although equally appreciable where mass innoculation isnecessary, for example in military scenarios, or in situations ofpandemic infection. For example, a typical single vaccine vial forpoultry vaccination comprises either 1000 or 2000 doses, and isregistered as such. Before freeze drying, a rough estimation is madeabout the titer of the material, but the final titer can only bedetermined after freeze drying, as the titer often decreases ratherunpredictably during freeze drying. Furthermore, the raw material oflive virus or bacteria cannot usually be kept stable long enough toobtain accurate titer results before freeze drying.

In practice, a container originally comprising more than 2000 doses canoften turn out to comprise only 1900 doses after freeze drying. In thatcase, the vial might only be marketable as a 1000 dose vial, since thattypically is the only other official registrable dosage. For such ascenario, a 47% waste of material and corresponding increase in costscould occur. This problem is compounded with combination vaccinesbecause the dosage of the component with the lowest titer must be usedto characterize the entire batch. As a further disadvantage, when vialsintended to provide 2000 single doses are subsequently marketed as 1000dose vials and used to innoculate 1000 animals, the animals thus dosedare unnecessarily over-exposed. Moreover, deliberately increasing thenumber of doses before freeze drying is not a satisfactory alternativewhen some of the vaccine batch is dried more efficiently. In thatsituation, one simply ends up wasting that material.

These problems will become increasingly important to solve asRegistration Authorities currently work toward a registration systemwhere the number of vaccine dose titers are set between well-definedupper and lower limits. Given the many variables in both the productionand the freeze drying process, it will be difficult to meet these limitson a large-scale production basis. As indicated, this problem will beeven more pronounced when a combination vaccine is required.

Yet a further disadvantage to previous freeze dried combination vaccinesis that a large number of formulated combinations must be kept instorage. This occurs because the various components in a combinationvaccine are typically mixed prior to freeze drying. Thus, for thepreparation of a full range of single/multi-component-vaccines against,e.g. two diseases, three different products must be kept in stock; (1)the product comprising anti-A vaccine, (2) the product comprising anti-Bvaccine; and (3) the product comprising anti-A and anti-B vaccine. Inthe case of vaccines against three diseases, seven differentvaccines/combinations have to be made and stored. For four diseases,this number mounts to fifteen different vaccines/combinations.Consequently, it is often necessary to provide and maintain a largestorage capacity.

U.S. Pat. Nos. 5,397,569 and 5,871,748 (Whitfill et al.) disclose amethod for producing active immunity against Newcastle Disease virus(NDV) in avian subjects by administering, in-ovo, a vaccine complexcomprised of a live vaccine virus and neutralizing antibodies boundthereto. Whitfill discloses that the ratio of virus to the neutralizingantibody or fragment thereof will determine the success of theimmunization. However, for such a method to be applied effectivelyagainst NDV, a narrow range of values for that ratio must be maintainedso that successful immunization will occur without killing the chicks.Moreover, the unpredictability in the titer using lyophilized NDV (aswith other virus preparations) means that methods such as Whitfill's arenot easily applicable for NDV. Therefore, a method which could ensuremore exact titers of NDV and antibodies for preparing such a vaccine isdesired.

Yet another disadvantage to prior vaccine formulations and their methodsof production lie in the space-consuming nature of the actual freezedrying process. Typically, lyophilizing a vaccine first requiresdispensing the solubilized vaccine into glass vials. The vials are thenloosely capped with rubber stoppers and placed on trays in racks in afreeze drying chamber or condenser. It is difficult to concentrate veryhigh doses of solubilized vaccine material in a very small volume.

As a result, the vials typically used in freeze drying for multiplevaccine doses always contain a relatively large volume of fluid.Efficient freeze drying requires that the fluid to be lyophilizedexposes a large surface area to the vacuum. Therefore, since only thetop of the frozen pellet is in contact with the vacuum, vaccines arefreeze dried in relatively large glass bottles or vials, with a widebottom. These vials are sometimes 5 centimeters high, wherein anadditional 2 cm of height is needed for the rubber stoppers.Consequently, the ratio of product material to empty space in the freezedrying apparatus is extremely inefficient leading to a less costeffective production process.

Another disadvantage to conventional compositions for freeze driedvaccines formulated to be contained in glass vials is that the rubberstoppers can hinder free transfer of water molecules from the vial tothe condenser during lyophilization. This can increase the partialpressure of sublimating gases inside the vial, thereby decreasing theefficiency of the lyophilization, as well as increase the risk ofcollapse of the lyophilized material. Glass also impedes heat transferto further affect lyophilization efficiency. Formulating a vaccine thatwould eliminate both glass vials and stoppers by using trays in thelyophilization would increase efficiency. Moreover, freeze drying isvery time-consuming. Vaccine components are typically kept just belowthe freezing point to decrease water sublimation time. However, a longerdrying period leads almost inevitably to a decrease of titer. Theglassvials with their stoppers and aluminum covers are generally seen asan encumbrance. In production terms they can represent more than 50percent of the cost of the finished vaccine. In field situations, thediluent must be injected into the vial and the resultant solutionextracted and diluted, if necessary for use. This is an inconveniencenot always suited to on-site situations, such as in a chicken-shed. Itis not uncommon for the operative in these situations to accidentally beselfinjected during this procedure. Yet another disadvantage to suchvaccine packaging and preparation is that some of the concentratedsolution will remain in the vial. Consequently, it is generally acceptedthat an overage in the contents is necessary to compensate fornon-homogeneity and production losses, as well as losses incurred overstorage periods. Glass vials must also be safely disposed of and canresult in possible health and environmental hazards.

As a result, use of conventional freeze dried vaccine formulationsinvolve complicated preparation techniques which are costly anddifficult to implement in the field. Consequently, such conventionalvaccine formulations are not well-suited for vaccinations inunderdeveloped countries were economic and field conditions do not allowfor costly campaigns, or in the case of mass immunizations which may beneeded, such as in the defense of biological warfare, or other potentialcatastrophes or epidemics.

What is desired, therefore, is a vaccine formulation which obviates theuse of glass vials for administration providing a less costly and bulkypackaging alternative. What is also desired is a vaccine formulation andmethod of use which provides greater dosing accuracy and ease of usewhile maintaining stability, sterility, solubility and homogeneity forboth single and multiple vaccine formulations. A vaccine formulation andimproved immunizing method which facilitates more accurate, reproducibleand efficient administration can provide the advantages of better fieldperformance, increased safety, cost-effectiveness, less waste, andimproved environmental compliance.

SUMMARY OF THE INVENTION

Accordingly, it is an object of the invention to provide a vaccinecomposition and method of immunizing accomplished by simply dissolving asolid, dense stable form of a lyophilized vaccine in a diluent therebyeliminating the need for glass vials, while also allowing for muchgreater accuracy of dosing and ease of use.

A further object of the invention is to provide a lyophilized live orinactivated vaccine which is compacted, compressed or tableted as adense stable solid that will retain its potential immunizing capacityduring preparation and for the duration required for sufficientshelf-life of a commercial vaccine.

A further object of the invention is to provide a lyophilized live orinactivated vaccine which is compacted, compressed or tableted as adense stable solid which facilitates formulation of in-ovo vaccinepreparations and uses thereof.

A further object of the invention is to provide a lyophilized live orinactivated vaccine which is compacted, compressed or tableted as adense stable solid which facilitates packaging of vaccines in non-vialcontainers and use thereof.

Another object of the invention is to provide a vaccine composition andmethod of immunizing with greater flexibility in the vaccinations thatcan be formulated and uses thereof.

A further object of the invention is to provide a vaccine compositionand method of immunizing which reduces the need for excess vaccinematerial needed to compensate for the inherent inaccuracies in the titerof the packaged vaccine made according to known methods.

Another object of the invention is to provide a vaccine composition andmethod of immunizing which allows greater flexibility in the types ofvaccine formulations that may be administered.

Yet another object of the invention is to provide a vaccine compositionand method of immunizing which faciliates formulation of a multi-dosevaccine solution and uses thereof.

Another object of the invention is to provide a vaccine composition andmethod of immunizing which facilitates human mass immunization.

Another object of the invention is to provide a vaccine composition andmethod of immunizing that is suitable and cost-effective forvaccinations in underdeveloped countries, where cost and fieldconditions do not allow for complicated preparation and costlycampaigns.

Yet another object of the invention is to provide a vaccine compositionand method of immunizing amenable for use in defense of biologicalwarfare, where mass immunization is a must.

These and other objects may be achieved by the present invention whichrelates to a stable vaccine composition comprising at least onepre-titrated lyophilized antigenic component and a dissolution aid,wherein the vaccine composition is in the form of a hard tablet. Inaddition, the present invention also provides a method of immunizing asubject against a disease comprising the steps of: dissolving a vaccinecomposition with a diluent to form a solution, wherein the vaccinecomposition comprises at least one lyophilized antigenic component and adissolution aid, wherein the vaccine composition is in the form of ahard tablet, and wherein the vaccine composition provides protectionagainst such disease; and administering the resulting solution to thesubject in an amount effective to immunize the subject against thedisease.

The invention in its particular features will become more apparent fromthe following detailed description considered with reference to theaccompanying examples. The following description will continue todiscuss the problems and solutions offered by the present invention asthey pertain to veterinary applications. However, the following shouldin no way be interpreted as being limited thereto or inapplicable tohuman vaccination.

DETAILED DESCRIPTION OF THE INVENTION

This invention provides a stable compressed vaccine compositioncomprising at least one lyophilized antigenic component and adissolution aid.

In a preferred embodiment of the invention, the vaccine composition isin the form of a hard tablet.

In another preferred embodiment of the invention, the compositiondissolves completely and rapidly in water.

In a preferred embodiment of the invention, the composition is sterile.

In a preferred embodiment of the invention, the dissolution aid is aneffervescent agent or pair of agents, a disintegrant, a sufactant, or asolubilizer.

In another preferred embodiment of the invention, the effervescent paircomprises a salt and an acid,

In another preferred embodiment of the invention, the effervescent pairis citric acid and the salt is a bicarbonate.

In another preferred embodiment of the invention, the vaccinecomposition comprises a binder and a lubricant.

In yet another preferred embodiment of the invention, the compositiondisintegrates completely and rapidly in water.

In another embodiment of the invention, the active material is apre-titrated lyophilizate that is kept under a dry nitrogen atmospherebelow a freezing temperature prior to use.

In another embodiment of the invention, the lyophilized componentcomprises up to 80% by weight of the composition.

In another preferred embodiment of the invention, the vaccinecomposition is characterized by complete dissolution within less than 80seconds upon contact with a diluent.

In another preferred embodiment of the invention, complete dissolutionof the vaccine composition occurs at a diluent to composition ratio ofbetween 0.5 ml of diluent per 200 mg of composition and 100 ml ofdiluent per 400 mg of composition.

In another preferred embodiment of the invention, the stability of thevaccine composition is characterized by a loss of titer no greater thanthe difference between 10^(9.7)EID₅₀/ml and 10^(9.2) EID₅₀/ml after 5days at 37° C.

In another preferred embodiment of the invention, the stability of thevaccine composition is characterized by a loss of titer no greater thanthe difference between 10^(9.7)EID₅₀/ml and 10^(9.3)EID₅₀/ml after 9months at 4° C.

In yet another preferred embodiment of the invention, the antigeniccomponent of the vaccine composition is a whole virus, a whole bacteriumor a whole microorganism.

In another preferred embodiment of the invention, the antigeniccomponent of the vaccine composition is a live bacterium or a livevirus.

In a preferred embodiment of the invention, the vaccine compositioncomprises an indicator of the presence of the antigenic component.

In a preferred embodiment of the invention, the indicator is a dye.

This invention further provides a stable vaccine composition comprisingat least one lyophilized antigenic component and a dissolution aid,wherein the vaccine composition is in the form of a hard tablet and theantigenic component comprises a lipopolysaccharide or a protein, whethernaturally occurring, recombinant, or modified.

In a another preferred embodiment of the invention, the antigeniccomponent is a live virus selected from the group consisting of:Newcastle Disease virus or bacterium, Newcastle Disease virus VH strain,Infectious Bursal Disease virus, Gumboro Disease virus, fowl pox virus,Laryngotraecheitis virus, Infectious Bronchitis of poultry virus,Gumboro Winterfield virus, Infectious Bronchitis (IB-H120), sheep poxvirus, Rinderpest virus, anthrax spores, Salmonella SPP., E. coli., oran admixture of one or more of the foregoing whether naturallyoccurring, recombinant or modified.

In yet another embodiment of the invention, the antigenic component isselected from the group consisting of: anthrax spores, Salmonella SPP,E. coli, or an admixture of one or more of the forgoing, whethernaturally occurring, recombinant or modified.

In yet another preferred embodiment of the invention, the antigeniccomponent is a live virus and the composition further comprisesneutralizing antibodies against the virus.

This invention further provides a method of immunizing a subject againsta disease comprising the steps of: dissolving a vaccine composition witha diluent to form a solution, wherein the vaccine composition comprisesat least one lyophilized antigenic component and a dissolution aid,wherein the vaccine composition is in the form of a hard tablet, andwherein the vaccine composition provides protection against suchdisease; and administering the resulting solution to the subject in anamount effective to immunize the subject against the disease.

In a preferred embodiment of the invention, the dissolving step of themethod of immunizing is further characterized by complete dissolution ofthe vaccine composition.

In yet another preferred embodiment of the invention, completedissolution during the dissolving step of the method of immunizingoccurs at a diluent to composition ratio of between 0.5 ml of diluentper 200 mg of composition and 100 ml of diluent per 400 mg ofcomposition.

In another preferred embodiment of the invention, the dissolving step ofthe method of immunizing is further characterized by completedissolution within less than 80 seconds upon contact with a diluent.

In another embodiment of the invention, the administering step comprisesinjecting the subject with the solution.

In another embodiment of the invention, the administering step comprisesspraying the subject with an aerosol formed from the solution.

In yet another embodiment of the invention, the administering stepcomprises applying the solution to the subject in the form of eye drops.

In yet another embodiment of the invention, the administering stepcomprises applying the solution by oral ingestion.

In a preferred embodiment of the invention, the method of immunizing isapplied to a subject that is an avian animal and the disease is selectedfrom the group consisting of Newcastle Disease, Infectious BursalDisease, fowl pox, Laryngotracheitis, Infectious Bronchitis of poultry.

In another preferred embodiment of the invention, the administering stepcomprises administering a solution to the subject in-ovo.

This invention also provides a package comprising the vaccinecomposition having at least one lyophilized antigenic component and adissolution aid, wherein the vaccine composition is in the form of ahard tablet or further comprises an indicator of the presence of theantigenic component.

In a preferred embodiment of the invention, the package is a blisterpack.

In another preferred embodiment of the invention, the package is asterile syringe.

This invention also provides a sterile syringe comprising a compactedvaccine composition, wherein the vaccine composition comprises at leastone lyophilized antigenic component and a dissolution aid.

In one embodiment of the invention, the compacted vaccine composition ofthe sterile syringe is a compressed composition.

In a preferred embodiment of the invention, the compressed vaccinecomposition of the sterile syringe is in the form of a hard tablet.

This invention further provides a method of immunizing a subject againsta disease which comprises adding a diluent to a sterile syringecomprising a compacted vaccine composition, wherein the vaccinecomposition comprises at least one lyophilized antigenic component and adissolution aid, or wherein the compacted vaccine composition of thesterile syringe is further characterized as a compressed composition, orwherein the compressed vaccine composition of the sterile syringe is inthe form of a hard tablet, so as to form a solution in the syringe andadministering the resulting solution to the subject.

Solutions resulting from dissolution of the vaccine composition of thepresent invention may be administered by any suitable means. The methodof immunizing a subject against a disease according to the presentinvention may employ a number of methods to administer a liquid solutionformed by the vaccine composition. Exemplary methods of administrationare intramuscular injection, subcutaneous injection, intravenousinjection, intra peritoneal injection, eye drop, via drinking water,aerosol, or nasal spray. When the animal to be treated is a bird, thebird may be a newly hatched (i.e., about one day old after hatching),adolescent, or adult bird. The vaccine of the present invention may beuseful for administration of birds in ovo, as described in U.S. Pat. No.4,458,630 to Sharma.

The freeze dried (lyophilized) pre-titrated vaccine composition of thepresent invention is understood to be a preparation composed of at leastone freeze dried antigenic pre-titrated vaccine component, excipientsand optionally various additives that have been compacted or compressedinto a dense form.

As used herein, “dense” refers to the vaccine composition having adensity greater than 1.0 g/cc. Usually, the density of the vaccinecomposition will be greater than 1.5 g/cc. Typically, the range ofdensity for the vaccine composition will be from about 1.5 g/cc to 2.5g/cc.

The vaccine composition may be compacted, compressed, or in the form ofa hard tablet.

As used herein, “compacted” refers to a vaccine composition having adensity greater than 1.0 g/cc, but no measurable hardness as measured inStrong-Cobb Units (SCU) and tested for hardness on a ERWEKA TabletHardness Tester Model TBH20.

As used herein, “compressed” refers to a vaccine composition having ahardness of at least 2.0 SCU and “hard tablet” refers to a vaccinecomposition in the form of a tablet or other dense form having ahardness of at least 3.0 SCU.

The compacted, compressed and hard tablets of the vaccine compositioncan be made on an instrumented MANESTY F3 Single Punch 12 mm FlatBeveled or 6 mm standard concave punches.

The vaccine composition in the form of a hard tablet was made at apressures of a maximum of 4 tonnes. The tablets were tested for hardnesson a ERWEKA Tablet Hardness Tester Model TBH20 as described above, andwere all found to have a hardness greater than 3.0 SCU.

The classic tablet normally associated with therapeutic agents isunderstood to be such a “tablet”. However, it is understood that anycompacted or compressed dense form is intended, including those havingless frequent use in the pharmaceutical field. For example, large“briquettes” would be suitable should the final application require alarge volume of material.

It is understood that a preparation that disintegrates or dissolves“completely” and “rapidly” in a diluent is one that is capable of doingso in pure water even if the intended use is not with pure water, butrather with a solution e.g., saline, or for that matter a non-aqueousvehicle.

By “completely dissolved” it is understood that no soluble component isleft undissolved. By “rapidly disintegrated or dissolved” is understoodthat disintegration or dissolution is complete within approximately afew minutes or less when a large volume of water is employed for smallvolume of compressed lyophilized vaccine composition e.g., 100 ml ofwater for a 400 mg effervescent tablet. The time is increased where thevolumes of diluent are comparably decreased. Thus the same tablet mightrequire 70 seconds with a volume of water of 10 ml, and 80 seconds in 2ml of water.

The disintegration or dissolution time referred to above is the timetaken for dissolution or disintegration of a tablet when placed in ameasured quantity of water at room temperature without stirring.

By “stable” it is understood that the compositions of the presentinvention will maintain their (potential) immunizing capacity duringpreparation and for the duration required for shelf life of a commercialvaccine.

As will be demonstrated by reference to the following examples, avaccine composition and method of immunization according to the presentinvention has numerous advantages as has been described herein.

The method of the present invention is generally exemplified as followsfor the preparation of a vaccine composition containing 1000 doses ofvaccine component. A solution of the vaccine component is lyophilized intrays. The dried material is pulverized. The titer of a homogeneouslymixed sample after freeze drying is determined while the lyophilizate iskept under nitrogen in a tightly secured container at −20° C. If thetiter of the lyophilized material is determined to be 625 doses per 100mg, it suffices to weigh out 160 mg of this material per compressedvaccine preparation to arrive exactly at the required titer. Thematerial is then mixed with optional and/or functional additives and thepreparation is compressed or tableted. The compressed freeze driedvaccine preparations can then be packaged by any of the methods known tothose skilled in the art and/or described herein.

The term excipient is a catch-all term for diluents or vehicles used inthe formulation of the vaccine composition. Excipients can include:diluents or fillers, binders or adhesives, dissolution aids, lubricants,antiadherents, glidants or flow promoters, colors, flavors, sweetenersand adsorbents.

Specifically, tablet fillers are substances that compromise the bulk ofthe tablet and primarily act as a carrier. Typical tablet fillersinclude, but are not limited to, calcium sulfate, calcium phosphate,calcium carbonate, starch, modified starches (carboxymethyl starch,etc.), microcrystalline cellulose, lactose, sucrose, dextrose mannitoland sorbitol.

Tablet filler levels are from about 0% to 90% by weight of the tablet.

Binders act as the “glue” which holds powders together to form granules.Binders include, but are not limited to, natural polymers such asstarches or gums acacia, tragacanth and gelatin or synthetic polymerssuch as PVP and methyl-, ethyl- and hydroxypropylcellulose.

Binder levels are from about 0% to 20% by weight of the composition.

Dissolution aids promote dissolution of the vaccine composition. Typicalexamples include, but are not limited to effervescent agents,disintegrates, surfactants and solubilizers.

Disintegrants cause compressed tablets to break apart. Typical examplesinclude, but are not limited to, starch, microcrystalline cellulose,purified wool starch, alginic acid, sodium starch glycolate guar gum,crosslinked polyvinyl pyrolidone (PVP), ion exchange resin andcelluloses such as methyl-, croscarmellose sodium, sodium carboxymethyl-and hydroxypropylmethyl-.

Dissolution aid levels are from about 1% to 95% by weight of thecomposition.

Lubricants reduce friction between the material to be compressed and diewall during compression and ejection. Most lubricants are waterinsoluble and include stearates (magnesium, calcium and sodium), stearicacid, talc and waxes. Water soluble lubricants include PEG's, sodiumbenzoate, sodium oleate, sodium acetate, sodium lauryl sulfate andmagnesium lauryl sulfate.

Lubricant levels are from about 0% to 5% by weight of the composition.

Colorants are added to help identify types of vaccine formulations suchas in the form of tablets for aesthetic and functional purposes, forexample and not as limitation to the present invention, the dyesdisclosed in Examples A through D taken from Israeli Patent No. 46189.

Colorant levels are from about <1% of the formulation.

The vaccine compositions of the present invention can be tabletedaccording to methods known to those skilled in the art, such asdescribed in Pharmaceutical Dosage Forms, Tablets, 2nd. Ed. 1989, Vols1, 2, 3, Editors H. A. Lieberman, L. Lachman, J. B. Schwartz.

In a preferred embodiment, the composition of the present invention is ahard tablet prepared having an effervescent agent as a dissolution aid.As those skilled in art appreciate, the effervescent tablet must containa basic component and an acidic component, such as an effervescent pair,so that upon dissolution appropriate reactions occur to generate carbondioxide and carbonic acid. Suitable effervescent components include thecarbonate family of basic compounds and inorganic or organic acidiccompounds.

Among the carbonate family of basic compounds, preferred effervescentagents for use in the compositions of the present invention are sodiumcarbonate, sodium bicarbonate, glycine carbonate, potassium carbonate,potassium bicarbonate, potassium dihydrogencitrate, and calciumcarbonate. A most preferred basic compound is sodium bicarbonate.

Preferred acidic components for use in the compositions of the presentinvention are citric acid, adipic acid, tartaric acid, maleic acid,boric acid, benzoic acid, hydroxybenzoic acid, methoxybenzoic acid,mandelic acid, malonic acid, lactic acid, pyruvic acid, glutaric acid,aspartic acid, hydrochloric acid, oxalic acid, salicylic acid, succinicacid, and acetic acid. A most preferred acidic effervescent component iscitric acid.

In addition to the basic and acidic effervescent tablet ingredientsdescribed above, the tablet composition of the present invention mayalso contain other excipients conventionally employed.

The tablet compositions of the present invention are obtained usingtableting procedures known in the art. Generally, the tabletingprocedures used for the present invention may be summarized as follows:the ingredients were sieved, blended and directly compressed intotablets of the required weight, size and hardness on a MANESTY F3 tabletpress.

The effervescent tablet compositions of the present invention may bedissolved in ordinary water or a simple saline solution.

Antigenic vaccine components are those components that specificallytrigger the immune response against the antigen or antigens from whichthe vaccine components were derived. Such components may originate fromone virus or microorganism including recombinants e.g. an antigeniclipopolysaccharide and an antigenic protein, or e.g., two differentantigenic proteins. They may also comprise antigenic parts of theprotein or polysaccharide. These components are generally referred to assubunit components. In some cases the vaccine component comprises thewhole virus or microorganism including recombinants. The vaccinecomponent can for example be a bacterin, or a live attenuated bacteriumor virus, an inactive bacterium or virus, or a dead bacterium or virusincluding recombinant and otherwise modified. Preferably, a vaccinecomponent is a live (modified nonvirulent or partially virulentincluding recombinant) bacterium or virus. Examples include, but are notlimited to, Salmonella bacteria, New Castle disease virus, InfectiousBursal Disease (Gumboro) virus, Infectious Bronchitis virus,Pseudorabies virus or anthrax spore vaccine.

Combination vaccines are vaccines comprising various vaccine components.Combination vaccines may also comprise antigenic components drived fromtwo or more different viruses or microorganisms.

More complex combinations are also possible. Thus, vaccines of one ofthe types described above, as well as mixtures thereof are referred toas combination vaccines. These can also contain any of the knownadjuvants.

A “package” is understood to be any package useful for the storage of astable compressed vaccine preparation.

The package may, for example and not as a limitation, be a glass orplastic (e.g., a high-density polyethylene) container generally used forpackaging and storage of tablets. Adding the stable compressed freezedried vaccine preparation to a diluent to homogeneously dissolve thepreparation would suffice to make the vaccine ready for use. Anotherform of packaging for compressed freeze dried vaccine preparations ofthe subject invention is a blister pack. One example of a blister packare plastic sheets, with rows of pits (trays) which contain thepreparations and are heat sealed with aluminum foil lidding preferablyunder dry nitrogen. Another variety of blister pack useful for sensitiveproducts are so-called aluminum/aluminum blisters where both tray andlidding are based on aluminum or aluminum laminates with variousplastics. This would avoid the use of expensive and space consumingglass vials, generally used for packaging and storage of vaccines forwhich the eventual user generally has no use.

The present invention makes it possible to add one or more compressedfreeze dried vaccine compositions directly from a blister pack to e.g.,a bucket of water to be used in the drinking water or aerosol sprayingsystem to ensure successful vaccination. This is usually accomplishedon-site, for example in a chicken shed. Veterinary vaccination can alsobe achieved by fogging, vent brush, piercing, whole body spray or as eyedrops. The vaccine solution obtained in a similar manner might be usedalternatively for in-ovo immunization of embryos as described in U.S.Pat. Nos. 5,397,569 and 5,871,748.

It is clear that any device that can be used to contain compressedsolids can be used in the invention. A diluent may be just water, or onthe other hand it may be a complex mixture of buffers and adjuvants.This will depend mainly on which additives have been added to thesolution prior to lyophilization or to the preparation prior tocompression.

Known freeze drying methods yield a cake comprising the vaccinecomponent or components in a vial. This method is space andtime-consuming, as mentioned. If the antigenic vaccine component orcomponents are freeze dried in trays of solution, then the whole surfaceof the cold plates in the freeze drying machine is utilized during theprocess. Additionally, should collapse occur, or for that matter ifthere is any other problem in the freeze drying process, the materialcan be easily recovered. This is not the case if the material waspre-filled into glass vials prior to lyophilization. Additionally, sinceno height consuming vials are involved in this part of the process, thecold plates can be stacked up to a high-density. As a result, thecapacity of freeze dryers substantially increases, limited practicallyonly by condenser capacity. As a corollary, much smaller freeze dryerscould be used for the present invention to achieve the same output aswhen vials are used in much larger dryers.

Therefore, in one preferred embodiment, the vaccine composition iscompressed from sterile lyophilized antigenic material which has beendried in trays and then broken into powder and admixed with sterileadditives to aid tableting and eventual dissolution. The sterile vaccinecomposition is then packaged under dry nitrogen gas in a sterile blisterstrip pack with a high moisture and gas barrier.

In one preferred embodiment, the lyophilized material is a liveattenuated virus.

In another preferred embodiment, an admix comprising a combination ofpreviously dried virus components is compressed into a singlepreparation together with additives to aid dissolution and optionallythose adjuvants desired for inoculation.

If a combination vaccine is required, the advantages offered by thepresent invention are even more pronounced. It suffices to simply addsufficient pre-titrated lyophilisate of each type of antigenic vaccinecomponent to the preparation before compression to achieve a combinationvaccine with each component in a highly accurate dose.

At the same time, the present invention offers a solution to the problemof the large storage capacity needed to store all possible variants ofe.g., a three or four component vaccine.

Instead of mixing the various components prior to freeze drying as maybe required for the freeze drying of combination vaccines, eachcomponent is freeze dried separately. Thus the various components can bestored separately. When necessary, each desired combination is easilymade by admixing the appropriate amount of lyophilized material of eachdesired component into one preparation as described herein, and thencompressing and packaging the mixture as described.

This allows for example, in the case of a 4-component combi-vaccine,stocking only four containers, each only containing lyophilized materialof one specific vaccine type to compose any single or combinationvaccine preparation that is required. Previous methods would require 15different containers, each comprising a prefabricated component ormixture.

In yet another preferred embodiment, the vaccine container comprises asterile blister strip containing solid forms, such as sterile tabletscompressed from lyophilized material of one or more vaccine componentsadmixed with a sterile dissolution aid in an effervescent pair (e.g.,sodium bicarbonate and citric acid) a sterile binder (e.g.,polyvinylpyrrolidone) and a sterile lubricant (e.g., sodium stearylfumarate, magnesium stearate). These single or multiple componentpreparations can then be used to easily prepare in situ an accuratevaccine solution of known titer. The foregoing are examples for thedissolution aid, binder and lubricant, and are not to be taken as alimitation for the vaccine composition of the present invention.

In yet another embodiment, the vaccine composition and method ofimmunization comprises a vaccine composition with components derivedfrom two or more antigenic components. A vaccine based on componentsfrom multiple antigenic components has the advantage that a singleadministration of such a vaccine suffices to induce protection againstmultiple diseases. It should be understood that for each virus ormicroorganism, any of several different antigenic vaccine components maybe included.

In yet a further embodiment, the antigenic components for each vaccinecan be mixed in the exact ratios needed to form a complex required forin-ovo immunization such as live Marek's disease, NDV and Gumborovaccines. Multicomponent in-ovo vaccine preparations are therefore alsopossible. The dosage form for such a combination can be achieved by thepresent invention by compressing an admixture of the freeze driedantigenic components for each disease, and the required antibodiestogether with the tableting aids. Such a preparation could then be usedto achieve the desired virus immunizing factor complex in situ bydissolution of the preparation in the required vehicle and understipulated conditions directly prior to in-ovo inoculation.

Usually, the vaccine container can contain between 1 and 10,000 doses ormore of vaccine. Single dose containers are common for individualvaccination, both for human and veterinary use.

Human Poliovirus vaccine, human live freeze dried Typhoid vaccine orCanine Parvovirus vaccine are typically formulated as single dosevaccines. It should be understood, that for a single dose of vaccine,similar formulation procedures as described for multi-dose vaccinesfully apply.

For the vaccination of large sheds of newly hatched chickens, it iscommon practice to use a 10,000 dose vaccine container for massvaccination against Infectious Bronchitis virus or NDV. For thevaccination of large farm animals such as cattle, against e.g.,Infectious Bovine Rhinotracheitis or Parainfluenza virus, vaccinecontainers with 10 doses are commonly used.

One of the advantages of the vaccine compositions prepared according tothe present invention, is that dissolution aids which may have beenincompatible in the freeze dried solution, can be included afterlyophilization. This speeds the administration procedure. It hasunexpectedly been found that the use of citric acid as part of aneffervescent pair, which was expected to be detrimental to the liveattenuated virus preparations, had no significant impact on the titers.

It should also be apparent that the integrity of the moisture barrierfor the container has a significant effect with formulations of thistype and steps should be taken to ensure that an effective moisturebarrier is formed in the packaging. This is particularly true whenemploying blister strip packaging and where the additives selected arepotentially destabilizing when wet, as is the case with citric acid.

The present invention also provides a method of immunizing a subjectagainst a disease comprising the steps of: dissolving a vaccinecomposition with a diluent to form a solution, wherein the vaccinecomposition comprises at least one lyophilized antigenic component and adissolution aid, wherein the vaccine composition is in the form of ahard tablet, and wherein the vaccine composition provides protectionagainst such disease; and administering the resulting solution to thesubject in an amount effective to immunize the subject against thedisease.

Alternatively, the lyophilisates may comprise antigenic vaccinecomponents which were derived from two or more viruses ormicroorganisms.

In addition, the present invention provides a vaccine compositionwherein the vaccine composition and a diluent are contained together ina package. Such an embodiment allows for a method of immunization wherea dissolving step to form a liquid vaccine for administration can occurin situ in the field.

It should be understood that a vaccine package or pack containing thecompressed freeze dried stable vaccine composition may be any usefulpackaging of a vaccine. In a simple form, the vaccine pack comprises avaccine container comprising the vaccine components, packed togetherwith instructions in a box. In a more complex form of a vaccine pack,the vaccine container could additionally comprise a diluent and asyringe. In an additional embodiment, the compressed lyophilized vaccineis packed under sterile conditions into a sealed syringe, where thedissolution of the vaccine can thus occur inside the sterile syringe. Inthis way the batch size of pre-filled syringes is not dependent on thespace occupied by syringes in the lyophilizer.

The following examples illustrate preparation and potency of the vaccinecomposition of the present invention when used to immunize a subjectagainst various infectious diseases. Stability evaluations with a titeranalysis of a compressed freeze dried tablet form for various vaccineformulations are also presented.

The examples are presented to further illustrate and explain the presentinvention and should not be taken as limiting in any regard. Unlessotherwise indicated in the examples and elsewhere in the specificationand claims, all parts and percentages are by weight. Temperatures are indegrees Centigrade.

EXAMPLE 1

1.1 Preparation of Tablets Comprising Live Attenuated Newcastle DiseaseVirus VH Strain (Hereinafter NDV-VH)

Fertile, specific-pathogen-free (SPF) eggs were inoculated with NDV andincubated according to known methods for propagation of NDV viruses ineggs.

Allantoic amniotic fluid (AAF) was harvested and purified bycentrifugation at 4000 g for 20 min.

Lyophilization in vials was performed fully according to known, standardprocedures.

Tablets were prepared in the following manner:

To 13.2 g of pulverized lyophilisate was added:

60 g sodium bicarbonate

42 g citric acid anhydrous

4.2 g polyvinylpyrrolidone K-30

0.3 g magnesium stearate

The ingredients were then mixed and tablets were punched each with aweight of 400 mg.

Each resultant tablet thus prepared contained 44 mg of the lyophilizate.

1.2 Analysis of Tablets

The titer in SPF eggs of a random sampling of the tablets was evaluatedand compared to the adjusted amount of the lyophilisate in the vials.

NDV-VH/tablet 10^(9.7) EID₅₀* NDV-VH/vial 10^(9.6) EID₅₀ *Egg infectiousdose 50 units.

When one tablet was added to 100 ml of water, dissolution was completein less than one minute, i.e. no trace of solid was visible to the nakedeye.

When one tablet was added to 10 ml of water, dissolution was complete in70 seconds.

When one tablet was added to 2 ml of water, dissolution was completeafter 80 seconds.

In 1 ml of water, although complete dissolution was achieved, theeffervescent nature of this tablet was not found to be involved in thefull dissolution, which had taken approximately 4 minutes.

1.3 Packaging and Stability

The tablets were packed in Polyvinyl chloride (PVC) blister strips andstability tested at 37° C. and found to be relatively stable over 5, 7and 9 day periods.

Days at zero 5 7 9 37° C. NDV-VH 10^(9.7) 10^(9.2) 10^(9.3) 10^(9.4)tablet

Storage at 4° C. produced no appreciable change in titer and color oftablets after nine months.

Months at 4° C. zero 3 9 NDV-VH tablet 10^(9.7) 10^(10.1) 10^(9.3)NDV-VH vial 10^(9.6)  10^(9.6) 10^(9.3)

Titers given are in EID₅₀ units per tablet or vial calculated for thesame amount of active material.

1.4 Potency Test of the Tableted NDV-VH in Chickens HI at 3 MortalityEID₅₀/ weeks Post- after Group NDV-VH Dose Vaccinaction challenge Atablet 1 10^(6.6) 9.3 2/18 B tablet 2 10^(6.3) 9.2 1/20 C vial 10^(6.6)9.2 2/19 D non- — 1.2 10/10  vaccinated HI = Haemagglutinationinhibition units.

A: A tablet of NDV-VH was dissolved in 20 ml of sterile distilled waterand 0.02 ml was used to vaccinate each of the 18 chicks (4 weeks old,kept in isolators) by eye drops.

B: Another group of chicks was similarly vaccinated with the tableted VHvaccine dissolved in 40 ml of sterile distilled water.

C: A vial of NDV-VH was dissolved in sterile distilled water to give thesame virus concentration as in A.

D: Non vaccinated control.

At three weeks post vaccination the birds were bled for antibodiestitration by the Haemagglutination inhibition (HI) test and all thegroups were challenged with viscerotropic velogenic NDV given by i.m.Injection at 10^(5.3) EID₅₀/bird at the Kimron Veterinary Institute (TheIsraeli Veterinary Services).

The results of the tests indicate that good protection is being providedto the chickens by all the vaccine preparations tested.

EXAMPLE 2

2.1 Preparation of Tablets Comprising NDV-VH

Fertile, SPF eggs were inoculated with NDV and incubated according toknown methods for propagation of NDV viruses in eggs.

AAF was harvested and purified by centrifugation at 4000 g for 20 min.Lyophilization was performed fully according to known, standardprocedures.

NDV-VH virus was lyophilized in a tray. The dried powder was sealedunder low humidity in a sterile double polypropylene bag and kept at−20° C. Effervescent tablets containing increasing contents of driedpowder were produced. Tablets were prepared according to the proceduredescribed in Example 1.

Composition Wt. (g) Wt. (g) Wt. (g) Wt. (g) VH 1.25 2.5 5.0 10.0Lyophilized (2.7%) (5.3%) (10.1%) (20.3%) tray dried Sodium 25.0 25.025.0 22.0 Bicarbonate Citric Acid 17.5 17.5 17.5 15.4 Anhydrouspolyvinyl- 1.75 1.75 1.75 1.75 pyrrolidone K-30 Magnesium 0.12 0.12 0.120.12 Stearate Tablet 0.5 0.45 0.38 0.36 weight

2.2 Analysis of Tablets

NDV-VH titers (EID₅₀) in the tablets were obtained by titrations in SPFeggs.

Percent  2.7  5.3 10.5 20.3 lyophilisate/ tablet NDV-VR 10^(8.00)10^(8.25) 10^(8.75) 10^(9.00) /tablet

Satisfactory linear relationship was obtained between active materialconcentrations and virus titers.

When one tablet was added to 100 ml of water, dissolution was completein less than one minute, i.e. no trace of solid was visible to the nakedeye.

2.3 Packaging and Stability

The tablets were placed in syringes, which were packed in aluminumlaminate sachets. An injectable solution was prepared by sucking thediluent for injection into the syringe, followed by complete dissolutionof the tablet. The resulting injectable solutions were used eitherdirectly, for individual vaccination, or alternatively for multiplevaccination after further dilution in a larger container. An automaticsyringe was then applied.

EXAMPLE 3

3.1 Preparation of Tablets Comprising NDV-VH Containing AntibodiesAgainst Newcastle Virus (NDA) for In-Ovo Vaccination

Fertile, SPF eggs were inoculated with NDV and incubated according toknown methods for propagation of NDV viruses in eggs.

AAF was harvested and centrifuged at 4000 g for 20 min. Lyophilizationwas performed fully according to known, standard procedures and thelyophilized material titrated. The desired amounts of NDV and NDA weredetermined.

Tablets were prepared in the following manner:

To 7.0 g of pulverized lyophilisate was added:

0.36 g lyophilized hyperimmune chicken serum for NDV (NDA)

13 g sodium bicarbonate

9.1 g citric acid anhydrous

0.9 g polyvinylpyrrolidone K-30

0.07 g magnesium stearate

The ingredients were then mixed and tablets were punched each with aweight of 500 mg. Each tablet thus prepared contained 115 mg of thelyophilized virus preparation.

3.2 Analysis of the Tablets

When one tablet was added to 100 ml of water, dissolution was completewithin 2.5 min, i.e. no trace of solid was visible to the naked eye.

3.3 Packaging

The tablets were packed in PVC blister strips.

3.4 Potency Test of Tableted NDV-VH with NDA In-Ovo

In-ovo vaccination of commercial broiler chicks with NDV-VH+NDA.Challenge of In-ovo vaccinated chicks at 2 and 4 weeks post hatching wasconducted. The embryos were injected in a commercial farm by anInovoject apparatus (Embrex U.S.A.) at 18 days of incubation. A tabletwas dissolved in 200 ml of Marek's vaccine diluent and 0.05 ml wasinjected into each egg. Both vaccinated and non-vaccinated chicks werekept in isolators to avoid non-specific contamination by NDV viruses.

HI at % Mortality % Mortality 2 at 2 weeks HI at 4 at 4 weeks weekschallenge weeks challenge Vaccinated 5.7 40 4.4 10 Non- 3.4 53 2.5 80vaccinated

At 2 and 4 weeks chicks were bled for antibodies detection by the HItest and both groups were challenged with viscerotropic velogenic NDVgiven by I.M. injection of 10^(5.3) EID₅₀/bird at the Kimron VeterinaryInstitute (The Israeli Veterinary Services). The results indicate thatgood protection was provided to the chicks at 4 weeks post hatching.

EXAMPLE 4

4.1 Preparation of Tablets Comprising Live Attenuated Gumboro DiseaseVirus (Hereinafter “MB”)

SPF eggs were inoculated with Gumboro (MB) disease virus and incubatedaccording to known methods for propagation of viruses in eggs.

Embryos and chorioallantoic membranes were harvested and the virus wasextracted by homogenization.

Lyophilization was done fully according to known, standard procedures.

Tablets were prepared in the following manner:

To 10.0 g of pulverized lyophilisate was added:

22.4 g sodium bicarbonate

15.7 g citric acid anhydrous

1.7 g polyvinylpyrrolidone K-30

0.1 g magnesium stearate

The ingredients were then mixed and tablets were punched each with aweight of 350 mg.

Each tablet thus prepared contained 70 mg of the lyophilisate.

4.2 Analysis of Tablets

The titer of a random sampling of the tablets was evaluated as 10^(5.6)EID₅₀/tablet.

(10^(2.5) EID₅₀ is considered to be a vaccinating dose)

When one tablet was added to 100 ml of water, dissolution was completein less than one minute, i.e. no trace of solid was visible to the nakedeye.

Virus titers in effervescent tablets and vials

MB/tablets 10^(5.6) EID₅₀ MB/vials* 10^(5.5) EID₅₀ *Adjusted to the samequantity of immunizing material.

4.3 Packaging and Stability

The tablets were packed in PVC blister strips and stability tested at37° C.

Days at 37° C. zero 5 MB 10^(5.6) 10^(4.9)

EXAMPLE 5

5.1 Preparation of Effervescent Tablets Containing Fowl Pox Live VaccineVirus

SPF eggs were inoculated with Fowl Pox disease virus and incubatedaccording to known methods for propagation of viruses in eggs.

Embryos and chorioallantoic membranes were harvested and the virus wasextracted by homogenization.

Lyophilization was performed fully according to known procedures.

700 mg tablets were prepared substantially as described in Example 1.

Each tablet contained 140 mg of lyophilisate.

5.2 Analysis of Tablets

The titer of each tablet was evaluated in SPF eggs and was found to be10^(6.25)EID₅₀/tablet. When one tablet was added to 100 ml of water,dissolution was complete in two minutes, i.e. no trace of solid wasvisible to the naked eye.

EXAMPLE 6

6.1 Preparation of Effervescent Tablets Containing Laryngotracheitis(Fowl) Live Virus

SPF eggs were inoculated with Laryngotrachietis vaccine virus andincubated according to known methods for growing viruses in eggs.Embryos and chorioallantoic membranes were harvested and the virus wasextracted by homogenization.

Lyophilization was performed fully according to known procedures. 700 mgTablets were prepared substantially as described in Example 1.

Each tablet contained 140 mg of lyophilizate. The titer of each tabletwas evaluated in SPF eggs and was found to be 10^(5.53) EID₅₀.

EXAMPLE 7

7.1 Preparation of Effervescent Tablets Containing Infectious Bronchitisof Poultry Live Virus

SPF eggs were inoculated with Infectious Bronchitis of Poultry diseasevirus and incubated according to known methods for growing viruses ineggs. Embryos and chorioallantoic membranes were harvested and the viruswas extracted by homogenization.

Lyophilization was performed fully according to known procedures.

700 mg tablets were prepared substantially as described in Example 1.

Each tablet contained 140 mg of lyophilisate.

7.2 Analysis of Tablets

The titer of each tablet was evaluated in SPF eggs and found to be10^(6.15) EID₅₀/tablet.

EXAMPLE 8

8.1 Preparation of Effervescent Tablets Containing Gumboro Winterfield(Fowl) Live Virus

SPF eggs were inoculated with Gumboro Winterfield (Fowl) disease virusand incubated according to known methods for propagating viruses ineggs.

Embryos and chorioallantoic membranes were harvested and the virus wasextracted by homogenization.

Lyophilization was performed fully according to known procedures.

700 mg tablets were prepared substantially as described in Example 1.

Each tablet contained 140 mg of lyophilizate.

8.2 Analysis of Tablets

The titer of each tablet was evaluated in SPF eggs and found to be10^(6.28) EID₅₀/tablet.

EXAMPLE 9

9.1 Preparation of Effervescent Tablets Containing Infectious Bronchitis(IB-HI20)+NDV-VH (Fowl) Live Virus

SPF eggs were inoculated with IB-HI20 or with NDV-VH disease virus andincubated according to known methods for propagating of viruses in eggs.

AAF was harvested and centrifuged at 4000/g for 20 min. Lyophilizationwas performed fully according to known procedures.

700 mg tablets were prepared as substantially described in Example 1.

Each tablet contained 140 mg of lyophilizate.

9.2 Analysis of Tablets

When one tablet was added to 100 ml of water, dissolution was completewithin 2 min, i.e. no trace of solid was visible to the naked eye.

The titer of each tablet was evaluated in SPF eggs and found to be10^(5.80) and 10^(9.60)EID₅₀/tablet for IB-HI20 and NDV-VH (Fowl)respectively.

EXAMPLE 10

10.1 Preparation of Effervescent Tablets Containing Sheep Pox Live Virus

Virus propagation was performed in Vero cells grown in plastic rollers.

Lyophilization was performed fully according to known procedures.

700 mg Tablets were prepared substantially as described in Example 1.

Each tablet contained 140 mg of lyophilizate

10.2 Analysis of Tablets

The titer of each tablet was evaluated as 10^(5.85) TCID₅₀/tablet. Sheeppox vaccine's virus was titrated in Vero cells

EXAMPLE 11

11.1 Preparation of Effervescent Tablets Containing Rinderpest LiveVaccine Virus (Cattle)

Virus propagation was performed in Vero cells grown in plastic rollers.

Lyophilization was performed fully according to known procedures.

700 mg Tablets were prepared substantially as described in Example 1.

Each tablet contained 140 mg of lyophilizate.

11.2 Analysis of Tablets

The titer of each tablet was evaluated as 10^(4.1) TCID₅₀/tablet.Rinderpest vaccine's virus was titrated in Vero cells.

11.3 Color Coding of Vaccine Compositions

Israeli Patent No. 46189 discloses poultry color-coded virus vaccinestained with a distinctive dye, which is safe in life tissues and whichdoes not harm the viability of viruses. The different vaccines are eachcolor-coded by means of a different distinctive dye. Suitable dyes are,for example, amaranth red dye, tartrazine yellow, indigo carmine blue ormixtures of any of these, resulting in distinctive hues.

A process of coloring the various different types of vaccines (orcolor-coding) of vaccines in different contrasting colors is usefulagainst mistakes that have been made by farmers who have vaccinatedtheir poultry flocks with a different type vaccine than the oneintended, thereby exposing their unprotected flocks against the diseasewhich they had actually intended to immunize. Such mistakes are usuallydiscovered only too late, if at all, and in various cases have broughtin complaints against the vaccine, in addition to serious financiallosses and damages to the farmer. Chemical colors, however, safe as theymay be by themselves, pose several problems and dangers when combinedwith a virus vaccine.

Vaccination of poultry involves the use of live virus vaccines in whicha culture of live viruses are contained in the final product, and sincelive viruses are very delicate and may deteriorate easily, the technicalproblem is quite involved. Different coloring agents and dyes are knownto be chemically active, namely they may oxygenate or reduce chemically.Live viruses are very sensitive to changes in the environment such as pHconditions, temperature, radiation, different metal ions and otherwisemay be affected by chemical agents with which they come in contact. Suchdyeing agents as methylene blue, or fuschin red or aniline dyes may betoxic to the viruses and possibly pose problems with the chickens orturkeys into which they would be injected. It was therefore necessary tochoose such dyes that will be safe for use in live tissues and will notharm the viability of the viruses with which they are in contact. Fourcolor shades have been chosen to identify the four most popular andimportant vaccines used in the poultry industry at present, namely a redcolor to identify Newcastle Disease vaccine of the lentogenic strain, oras it is called in Israel, the Komarov “chick” strain; a yellow color toidentify the Newcastle Disease vaccine of the Komarov-Haifa or “pullet”strain; a blue color to identify the Laryngotracheitis vaccine; and agreen shade to identify the Fowl Pox vaccine.

As disclosed in Israeli Patent No. 46189, different concentrations ofthese dyes were tried and were mixed directly with the wet vaccinebefore it was dispensed into the final containers and before it wasdesiccated (lyophilized). Several tests were run to ascertain that thesedyes will not harm the vaccine.

Different batches of dyed vaccines were subjected to storage conditionsunder refrigeration and otherwise and repeatedly titrated to verify thevirus concentration. Ample controls of the vaccine without the additionof the dyes were run parallel to every test done on the dyed vaccine.Dyed vaccines have also been tested directly on live birds to see whateffect they might have. Repeat tests with all four types of vaccines andfour dyes used in these experiments have shown no detrimental effect onthe virus titration results. Injection trials in live birds and theapplication of the virus vaccines by other routes such as through thedrinking water; by the stick method through the skin (Fowl Pox); and bythe vent-brush method (for laryngothracheitis vaccine) have shown thatthe dye had no detrimental effect on the birds and when injectedintramuscularly it was reabsorbed from tissues and completelydisappeared within 36 hours. Israeli Patent No. 46189 discloses vaccinestested in the field on several flocks which were vaccinated from one dayof age and up to marketing age with these dyed vaccines. These birdswere successfully marketed and had no evidence of the dyes whatsoever.

Further details of the invention disclosed in Israeli Patent No. 46189may be readily understood in connection with the description given inthe following examples taken therefrom, and which have been selected forthe purpose of illustration only in order to enable one skilled in theart to use the color-coded formulations for vaccines, and thus are notlimitations upon the invention.

Example A

AMARANTH red dye was prepared for combination with Vineland Newcastle“chick” vaccine, by making a concentrate-dye-solution. This concentratecontained 145 grams of dry powdered dye dissolved in 4 liters ofdistilled water. The concentrated dye solution was autoclaved at 20 psifor 20 minutes shortly after preparation and was stored in sealed glassbottles in a dark place. The dye-concentrate solution was added at 1.33%to the final Vineland Newcastle-Chick-Vaccine.

Results of the titrations with dyed vaccine compared to undyedcontrolled have been found to be as follows:

Titrations 2 weeks after lyphilization: without dye 10^(9.0) EID 50/mlwith dye 10^(9.3) EID 50/ml Accelerated titration: without dye 10^(9.1)EID 50/ml with dye 10^(9.3) EID 50/ml Titration 30 days afterlyophilization: without dye 10^(9.16) EID 50/ml with dye 10^(9.3) EID50/ml Titration 3 months after lyophilization: without dye 10^(8.86) EID50/ml with dye 10^(9.3) EID 50/ml Titration done 6 months afterlyophilization only sample 10^(9.16) EID 50/ml with dye

Example B

TARTRAZINE yellow dye was prepared for combination with VinelandNewcastle Pullet Strain Vaccine (The Mesogenic Haifa-Komorav Strain), bymaking a concentrate-dye-solution. This concentrate contained 200 gramsof dry powdered dye dissolved in 3 liters of distilled water. Theconcentrated-dye-solution was autoclaved at 20 psi for 20 minutesshortly after preparation and was stored in sealed glass bottles in adark place. The dye-concentrate solution was added at 3% to the finalVineland Newcastle Pullet Vaccine.

Results of the titrations with dyed vaccine compared to undyed controlshave been found to be as follows:

Results of regular vaccine titration: 10^(9.4) EID₅₀ per milliliterResults of titrated sample with dye: 10^(8.8) Titration after 7 daysincubation at 37° C.: without dye 10^(8.3) EID 50/ml with dye 10^(8.8)EID 50/ml Titration 39 days after lyophilization: without dye 10^(8.66)EID 50/ml with dye 10^(8.9) EID 50/ml Titration 93 days afterlyophilization: without dye 10^(8.66) EID 50/ml with dye 10^(8.66) EID50/ml 7 ½ months after lyophilization: without dye 10^(8.53) EID 50/mlwith dye 10^(8.76) EID 50/ml

Example C

INDIGO-CARMINE blue dye was prepared for combination withVineland-Injections Laryngotracheitis Vaccine, by making aconcentrate-dye-solution.

This concentrate contained 200 grams of dry-powdered dye dissolved in 3liters of distilled water. The concentrate dye solution was autoclavedat 20 psi for 20 minutes shortly after preparation and was stored insealed glass bottles in a dark place. The dye concentrate solution wasadded at 3% to the final Vineland Injections Laryngotracheitis Vaccine.

Results of the titrations with dyed vaccine compared to undyed controlshave been found to be as follows:

Titration 3 weeks after lyophilization: without dye 10^(6.3) EID 50/mlwith dye 10^(6.16) EID 50/ml Accelerated titration: without dye 10^(5.0)EID 50/ml with dye 10^(4.8) EID 50/ml Titration 30 days afterlyophilization: without dye 10^(6.0) EID 50/ml with dye 10^(5.8) EID50/ml Titration 3 months after lyophilization: without dye 10^(5.66) EID50/ml with dye 10^(5.5) EID 50/ml Titration 7 months afterlyophilization: without dye 10^(5.5) EID 50/ml with dye 10^(5.6) EID50/ml

(This is a somewhat unusual drop in titre for both dyed and undyedsamples and accelerated batched showed more that 1 log drop onacceleration indication possible high moisture content in this batch.)

Example D

A green shade dye coded “P” (for Pox) was obtained by mixingINDIGO-CARMINE with TARTRAZINE dyes and was used for combination withVineland Fowl Pox Vaccine. 100 grams of powered dye mix was dissolved in3 liters of distilled water.

Same procedure was used as in examples No. 2 and No. 3 and the solutionwas added at 3% to the final Fowl Pox vaccine.

Results of the titrations with dyed vaccine compared to undyed controlshave been found to be as follows:

7 days after lyophilization undyed control: 10^(5.5) EID 50/ml vaccinewith dye: 10^(5.5) EID 50/ml 3 months later undyed control: 10^(5.4) EID50/ml vaccine with dye: 10^(5.5) EID 50/ml 6 months later undyedcontrol: 10^(5.35) EID 50/ml vaccine with dye: 10^(5.4) EID 50/ml 9months after production undyed control: 10^(3.1) EID 50/ml vaccine withdye: 10^(5.3) EID 50/ml

SUMMARY OF THE INVENTION

(continued)

A further object of the invention is to provide a sterile packagecomprising a syringe containing a formulation which is compacted,compressed or tableted as a dense solid having a dissolution aid.

A further object of the invention is to provide the above sterilepackage, wherein the compacted, compressed or tableted solid formulationdissolves rapidly in a diluent, either within the syringe or in aseparate container.

Another object of the invention is to provide a vaccine composition andmethod of immunizing accomplished by dissolving a solid, dense stableform of a lyophilized vaccine contained in a sterile package with adiluent to form a vaccine solution ready for administration.

A further object of the invention is to provide a lyophilized live orinactivated vaccine which is compacted, compressed or tableted as adense, stable, solid suitable for human administration.

Another object of the invention is to provide a lyophilized live orinactivated vaccine which is compacted, compressed or tableted as adense, stable, solid further comprising a second lyophilized componentcontaining neutralizing antibodies against a lyophilized antigeniccomponent.

A further object of the invention is to provide a process for producingthe lyophilized vaccine containing neutralizing antibodies where thepotency of the lyophilized antigenic component and the lyophilizedneutralizing antibodies are separately determined to produce a vaccineformulation in a ratio for each component based on their determinedpotencies.

Yet another object of the invention is to provide a method of immunizinga subject in-ovo accomplished by simply dissolving a solid form of thelyophilized vaccine containing neutralizing antibodies either within apre-packaged sterile syringe or in a separate container.

DETAILED DESCRIPTION OF THE INVENTION

(continued)

This invention further provides a sterile syringe containing a stable,compressed composition, wherein the compressed composition comprises adissolution aid.

In another embodiment, the invention provides the sterile syringecontaining a stable, compressed composition having a dissolution aid,wherein the compressed composition is in the form of a hard tablet.

In a preferred embodiment, the invention provides the sterile syringecontaining the hard tablet described above, wherein the hard tabletdissolves completely and rapidly in a diluent.

In a another preferred embodiment, the invention provides the sterilesyringe containing the hard tablet described above, wherein thecompressed composition is selected from the group consisting of aninjectable hormone, drug, and vitamin.

In another embodiment, the invention provides a method of administeringan injectable solution to a subject which comprises adding a diluent toany of the syringes described above to form an injectable solution inthe syringe and administering the resulting solution to the subject.

This invention also provides a package comprising a stable compressedvaccine composition comprising at least one lyophilized antigeniccomponent and a dissolution aid, wherein the package comprises a vial.

In a preferred embodiment of the invention, the vial is evacuated.

In another preferred embodiment of the invention, the vial is glass.

In a preferred embodiment of the invention, the package is a sachet.

This invention also provides a method of immunizing a subject against adisease comprising the steps of dissolving the vaccine composition whichprovides protection against such disease with a diluent to form asolution; and administering the resulting solution to the subject in anamount effective to immunize the subject against the disease, whereinthe subject is a mammal.

In a preferred embodiment of the invention, the mammal is selected fromthe group consisting of a human subject, a bovine subject, and an ovinesubject.

In a preferred embodiment of the invention, the antigenic component ofthe vaccine composition is a tetanus toxoid.

In a preferred embodiment of the invention, the mammal is a humansubject and the lyophilized antigenic component of the vaccinecomposition is a tetanus toxoid.

This invention further provides a stable compressed vaccine compositioncomprising at least one lyophilized antigenic component and adissolution aid and a second lyophilized component, wherein the secondlyophilized component contains neutralizing antibodies against thelyophilized antigenic component.

In a preferred embodiment, the invention provides the stable compressedvaccine composition having the second lyophilized component containingneutralizing antibodies, wherein the antigenic component is a live virusselected from the group consisting of: Newcastle Disease virus,Infectious Bursal Disease virus, fowl pox virus, Laryngotracheitisvirus, Infectious Bronchitis of poultry virus, sheep pox virus,Rinderpest virus, or an admixture of one or more of the foregoing,whether naturally occurring, recombinant or modified.

In another embodiment, the invention provides the stable compressedvaccine composition having a second lyophilized component containingneutralizing antibodies produced by the process comprising the steps of(a) lyophilizing at least one antigenic component; (b) lyophilizingneutralizing antibodies against the antigenic component separately fromthe antigenic component; (c) determining the potency of each of thelyophilized antigenic component and the lyophilized neutralizingantibodies; (d) mixing the lyophilized antigenic component and thelyophilized neutralizing antibodies in a ratio determined based on thepotencies obtained; and (e) compressing the mixture of the lyophilizedantigenic component and the lyophilized neutralizing antibodies anddissolution aids to form a stable compressed vaccine composition.

In yet another embodiment, the invention provides a method of making astable compressed in-ovo vaccine composition comprising the steps of (a)lyophilizing at least one antigenic component; (b) lyophilizingneutralizing antibodies against the antigenic component separately fromthe antigenic component; (c) determining the potency of each of thelyophilized antigenic component and the lyophilized neutralizingantibodies; (d) mixing the lyophilized antigenic component and thelyophilized neutralizing antibodies in a ratio determined based on thepotencies obtained, and (e) compressing the mixture of the lyophilizedantigenic component and the lyophilized neutralizing antibodies anddissolution aids to form a stable compressed in-ovo vaccine composition.

In a preferred embodiment, the invention provides the above method ofmaking a stable compressed in-ovo vaccine composition, wherein theantigenic component is a live virus selected from the group consistingof: Newcastle Disease virus, Infectious Bursal Disease virus, fowl poxvirus, Laryngotracheitis virus, Infectious Bronchitis of poultry virus,Marek's Disease virus, coccidiosis or an admixture of one or more of theforegoing, whether naturally occurring, recombinant or modified.

In another embodiment, the invention provides a method of immunizing asubject in-ovo against a disease comprising the steps of (a) dissolvingthe stable compressed vaccine composition providing protection againstthe disease having the second lyophilized component containingneutralizing antibodies with a diluent to form a solution; and (b)administering the resulting solution to the subject in-ovo in an amounteffective to immunize the subject against the disease with no adversereaction.

In a preferred embodiment of the above method of immunizing a subjectin-ovo, the subject is an avian subject and the disease is selected fromthe group consisting of Newcastle Disease, Infectious Bursal Disease,fowl pox, Laryngotracheitis, Marek's Disease virus and InfectiousBronchitis of poultry.

Further examples are presented for the preparation of human and animaltetanus toxoid vaccine compositions according to the present invention.

EXAMPLE 12

12.1 Preparation of 100 Dose Tablets Containing Tetanus Toxoid Vaccine

A liquid solution of Tetanus toxoid (Pasteur Merieux Connaught, Lot.-JJ,4750 LF units/ml) was lyophilized in vials. The lyophilized powder wascalculated to contain 47,500 LF units/gram.

One human immunization dose is 4 LF units (Micromedex Inc. HealthcareSeries Vol. 107).

100 dose tablets are prepared with a single punch tableting machine suchas an instrumented MANESTY F3 Single Punch 6 mm standard concave punchesaccording to the following formulation.

Formulation:

TABLE 12 Tetanus toxoid lyophilized powder 1.85 g NaHCO₃ anhydrous(Merck, 22.9 g Darmstadt Germany) Citric acid (Gadot, Israel) dry 14.0 gPovidone (PVP) 1.4 g Magnesium stearate 0.1 g

The dry ingredients, excluding the sterile tetanus toxoid, weresterilized by gamma irradiation.

The preparation tools were sterilized by steam at 121° C. and 15 PSI for45 minutes.

The tableting powders were passed through 30 mesh and thoroughly mixedin a polypropylene bag.

Tablets of 0.2 grams were compressed by the tableting machine to ahardness of at least 2.0 SCU and immediately packed in a 10 ml sterilesyringe. The syringe was immediately packed in a laminated aluminumsachet under a stream of dry, sterile nitrogen gas. The sachet wassealed and stored at 4° C.

Dissolution of the tablet was examined by opening the sachet, sucking 5ml of sterile reverse osmosis (RO) water at a temperature of 20° C. intothe syringe and pulling the piston of the syringe all the way up, theneedle directed upwards to allow the gas produced in the effervescentprocess to be released. After the tablet was completely dissolved sothat a clear suspension could be observed, the piston was pushed down tothe suspension front. Dissolution time was 35 sec. with 5 mls of ROwater at a temperature of 20° C. The liquid was transferred asepticallyto 45 ml RO water giving a final volume of 50 ml.

EXAMPLE 13

13.1 Preparation of 10 Dose Tablets Containing Tetanus Toxoid Vaccine

A liquid solution of Tetanus toxoid (Pasteur Merieux Connaught, Lot.-JJ,4750 LF units/ml) was lyophilized in vials and harvested in a drysterile environment into a homogeneous pool. The lyophilized powder wascalculated to contain 47,500 LF units/gram.

One human immunization dose is 4 LF units (Micromedex Inc. HealthcareSeries Vol. 107).

10 dose tablets are prepared with a single punch tableting machine suchas an instrumented MANESTY F3 Single Punch 6 mm standard concave punchesaccording to the following formulation.

Formulation:

TABLE 13 Tetanus toxoid lyophilized 0.185 g powder NaHCO₃ anhydrous(Merck, 24.5 g Darmstadt Germany) Citric acid (Gadot, Israel) 14.0 g dryPovidone (PVP) 1.4 g Magnesium stearate 0.1 g

The dry ingredients, excluding the sterile tetanus toxoid, weresterilized by gamma irradiation.

The preparation tools were sterilized by steam at 121° C. and 15 PSI for45 minutes.

The tableting powders were passed through 30 mesh and thoroughly mixedin a polypropylene bag.

Tablets of 0.2 grams were compressed by the tableting machine to ahardness of at least 2.0 SCU and immediately packed in a 10 ml sterilesyringe. The syringe was immediately packed in a laminated aluminumsachet under a stream of dry sterile nitrogen gas. The sachet was sealedand stored at 4° C. Dissolution of the tablet was examined by openingthe sachet, sucking 5 ml of sterile RO water at a temperature of 20° C.into the syringe and pulling the leading pole of the syringe all the wayup, the needle directed upwards to allow the gas produced in theeffervescent process to be released. After the tablet was completelydissolved so that a clear suspension could be observed, the piston waspushed down to the suspension front.

Dissolution time was 37 sec. with 5 mls of RO water at a temperature of20° C.

EXAMPLE 14

14.1 Preparation of Single Dose Tablets Containing Tetanus ToxoidVaccine

A liquid solution of Tetanus toxoid (Pasteur Merieux Connaught, Lot.-JJ,4750 LF units/ml) was lyophilized in vials and harvested in a drysterile environment into a homogeneous pool. The lyophilized powder wascalculated to contain 47,500 LF units/gram.

One human immunization dose is 4 LF units (Micromedex Inc. HealthcareSeries Vol. 107).

Single dose tablets are prepared with a single punch tableting machinesuch as an instrumented MANESTY F3 Single Punch 6 mm standard concavepunches according to the following formulation.

Formulation:

TABLE 14 Tetanus toxoid lyophilized 0.0185 gr powder NaHCO₃ anhydrous(Merck, 24.7 gr Darmstadt Germany) Citric acid (Cadot, Israel) dry 14.0gr Povidone (PVP) 1.4 gr Magnesium stearate 0.1 gr

The dry ingredients, excluding the sterile tetanus toxoid, weresterilized by gamma irradiation.

The preparation tools were sterilized by steam at 121° C. and 15 PSI for45 minutes.

The tableting powders were passed through 30 mesh and thoroughly mixedin a polypropylene bag.

Tablets of 0.2 grams were compressed by the tableting machine to ahardness of at least 2.0 SCU and immediately packed in a 5 ml sterilesyringe. The syringe was immediately packed in a laminated aluminumsachet under a stream of dry sterile nitrogen gas. The sachet was sealedand stored at 4° C.

Dissolution of the tablet was examined by opening the sachet, sucking0.5 ml of sterile reverse osmosis (RO) water at a temperature of 20° C.into the syringe and pulling the piston of the syringe all the way up,the needle directed upwards to allow the gas produced in theeffervescent process to be released.

After the tablet was completely dissolved so that a clear suspensioncould be observed, the piston was pushed down to the suspension front.

Dissolution time was 38 sec. with 0.5 mls of RO water at a temperatureof 20° C.

As a result of the stable, compressed vaccine composition according tothe present invention, a sterile package, such as a syringe, containingthe vaccine tablet can be readily used in the field, thereby eliminatingextra steps and materials needed to prepare the vaccine and vaccinate asubject.

The vaccine composition of the present Invention provides the advantageof facilitating preparation and administration of the vaccine becauseits formulation allows for packaging under sterile conditions into e.g.,a sterile syringe, where its dissolution can be accomplished usingstandard handling procedures for injectables. Consequently, once asterile vaccine tablet according to the present invention is packed in asterile syringe, a complete sterile route can be provided frommanufacturing to administration. This provides a marked advantage overthe method of packaging the tablets in blisters, where the tablet isexposed to a non-sterile environment upon contact with a diluent.

Vaccine tablets of single, as well as multiple doses can be efficientlyhandled in this technique.

The vaccine composition of the present invention can readily form avaccine suitable for parenteral administration, which allows the vaccinecomposition to be not only a live vaccine, but also an inactivated one.For example, as stated above, dead vaccine components can be either, aninactivated whole cell or virus, a subunit vaccine, either recombinantor a non-genetically modified one, or a toxoid.

The above suggested methods are suitable for animal as well as humanprophylaxis.

In addition, packaging of the vaccine composition of the presentinvention, for example, in the form of a tablet in a sterile package,e.g. a syringe, for storage and subsequent dissolution andadministration by injection is not only suitable for vaccinationmaterial. This technique can be used for other injectables such ashormones, drugs, vitamins, etc.

The methods described in the specification to produce a stable,compressed composition contained in a sterile package provide thefollowing manufacturing benefits: the active material is accuratelyexamined/analyzed prior to compression or tableting, the distribution ofthe tablets into, for example, open syringe cylinders is easilyaccomplished in a continuous process and many materials can becompressed into the tablet in a compacted volume. Lyophilized materialscan be included in the tablet and can keep their characteristics asshown in the above examples. Moreover, a dissolution aid is added to thecomposition, such as in the case of effervescent tablets to allow forcomplete and/or rapid dissolution with no major effect on the activematerials.

The syringes filled with the tablets can be packed into a sachet underdry, sterile nitrogen gas. This ensures a prolonged shelf life of theproduct.

The above description is for the purpose of teaching the person ofordinary skill in the art how to practice the invention, and is notintended to detail all those obvious modifications and variations of it,which will become apparent to the skilled worker upon reading thedescription. It is intended, however, that all such obviousmodifications and variations be included within the scope of the presentinvention, which is defined by the following claims.

What is claimed is:
 1. A sterile syringe containing a stable, compressedcomposition, wherein the compressed composition comprises a dissolutionaid, and at least one component selected from the group consisting of anantigenic component, an injectable hormone, a drug, and a vitamin. 2.The sterile syringe of claim 1, wherein the compressed composition is inthe form of a hard tablet.
 3. The sterile syringe of claim 2, whereinthe hard tablet dissolves completely and rapidly in a diluent.
 4. Amethod of administering an injectable solution to a subject whichcomprises adding a diluent to the syringe of claim 1 to form aninjectable solution in the syringe and administering the resultingsolution to the subject.
 5. A method of administering an injectablesolution to a subject which comprises adding a diluent to the syringe ofclaim 2 to form an injectable solution in the syringe and administeringthe resulting solution to the subject.
 6. A stable compressed vaccinecomposition comprising at least one lyophilized antigenic component anda dissolution aid, wherein the antigenic component is a tetanus toxoid.